This invention relates to an improved method for the manufacture of ethanol from cellulose. By the process of this invention a cellulose-containing substrate is reacted in one step with cellulase enzymes and with a means to manufacture ethanol.
The manufacture of ethanol from cellulose and cellulose-containing materials in one step has been disclosed and claimed in U.S. Pat. No. 3,990,944, Gauss, Suzuki, Takagi. Variations of the method have been claimed by Hoge in U.S. Pat. No. 4,009,075 and others.
Additionally, Metzger in U.S. Pat. No. 3,711,392 and Lang in U.S. Pat. No. 4,094,740 and U.S. Pat. No. 4,093,516 disclose the manufacture of ethanol from organic waste material.
Hoge discloses the use of Saccharomyces cerevisiae as the yeast useful for converting the glucose produced in situ into ethanol. Gauss et al. discloses the use of Saccharomyces cerevisiae, and Rhizopus javanicus for the same purpose. Neither of the Lang patents nor Metzger provide a disclosure of specific yeasts which will convert glucose to ethanol. Other yeasts which are well-known for their ability to convert glucose to ethanol include Baker's yeast and Saccharomyces carlsbergensis.
Amano, et al., "A Strongly Ethanol Assimilating New Yeast", The Society of Fermentation Technology, Japan, 53, 311 (1975) discloses the yeast Candida brassicae, IFO 1664, ATCC 32196, and its unique and unobvious differences from other well-known Candida species. According to Amano, et al., the following constitutes a description of Candida brassicae, ATCC 32196.
Cultura in extracto malti: add 25.degree. C., post 3 dies cellulae ovoideae ad cylindricae, 2.5-5.times.3-17.5.mu., singulares vel catenatae; pseudomycelium praesens. Sedimentum et pellicula formantur.
Cultura in striis agaro malti: add 17.degree. C., post unum mensem color albidus ad flavalbidus, pagina opaca, elevata, mollis et laevis vel crispulata. Pseudomycelium praesens abundanter.
Ascosporae, ballistosporae et teliosporae nullae.
Fermentatio: glucosum.
Assimilatio originum carbinum: glucosum, galactosum, sucrosum, maltosum (valde exigue), raffinosum (exigue, D-xylosum (exigue), ethanolum, glycerolum, acidum lacticum et acidum succinicum.
Kalii nitras non assimilatur, arbutinum non finditur, ureum non finditur. Vitamina addita non necessaria sunt.
Temperatura maxima crescentiae 43.degree.-45.degree. C.
TYPUS: culture RIFY E-17 in collectione Research Institute of Fermentation, Yamanashi University, Kofu, isolata e flio Brassica oleracea var. capitata in Kofu, Japonia, 1973.
Growth in malt extract: After 3 days at 25.degree. C., cells are short-oval to cylindrical, (2.5-5).times.(3-17.5).mu., single, in pairs or in chains (FIG. 1). Pseudomycelical cells are formed (FIG. 2). A sediment and thin wrinkled pellicle are formed.
Growth on malt agar: After one month at 17.degree. C., the streak culture is white to slightly yellowish, dull, raised, soft and smooth or slightly wrinkled.
Slide cultures on potato agar: A pseudomycelium with a tree-like formation is abundantly formed (FIGS. 3 and 4).
Formation of spores: No ascospore, ballistospore or teliospore is formed:
Fermentation:
______________________________________ Fermentation: Glucose + Trehalose - Galactose - Lactose - Sucrose - Raffinose - Maltose - Xylose - Assimilation of carbon compounds: Glucose + D-Ribose - Galactose + L-Rhamnose - L-Sorbose - Ethanol + Sucrose + Glycerol + Maltose vw Erythritol - Cellobiose - Ribitol - Trehalose - Galactitol - Lactose - D-Mannitol - Melibiose - D-Glucitol - Raffinose w .alpha.-Methyl-D-glucoside - Melezitose - Salicin - Inulin - DL-Lactic acid + Soluble starch - Succinic acid + D-Xylose w Citric acid - L-Arabinose - Inositol - D-Arabinose - ______________________________________ Assimilation of potassium nitrate: Absent. Growth in a vitaminfree medium: Good growth. Sodium chloride tolerance: 14-15% (w/v). Maximum temperature for growth: 43-45.degree. C. Splitting of arbutin: Negative. Hydrolysis of urea: Negative.